Building Better Antioxidants: Virtual Screening, Synthesis, and Characterization of Multifunctional Small Molecules Combining Nrf2 Pathway Activation and Direct Antioxidant Activity

项目总结/摘要 不受控制的氧化应激有助于神经退行性疾病和眼部疾病的发展， 目前还没有有效的治疗干预措施。调查人员已经开发出了小 在生物测定中通过两种机制表现出显著抗氧化反应性的分子：直接 与活性氧的反应性和Nrf 2途径的催化活化。调查人员将 结合虚拟筛选、合理设计、合成化学和分子生物学方法， 生产增强的抗氧化剂小分子，并了解小分子如何激活催化剂， 抗氧化途径 该项目产生的新分子有望提供自由基清除活性，代谢 稳定性和渗透性。我们提出了四个机制，实现这些功能的改善 与母体分子相比。虚拟筛选将用于确定这些组合 这些方法将产生最有希望的目标。在合成这些靶标之后， 神经退行性疾病和眼部疾病（例如白内障）将用于测试对氧化应激的保护。 显示出有效抗氧化活性的分子随后将被筛选用于代谢稳定性和血脑 屏障渗透性探索还将侧重于揭示我们的 小分子通过生物学中先天抗氧化剂途径的催化活化来保护（Nrf 2）。细胞 将评价来自暴露于ROS的模型的裂解物的Nrf 2及其下游基因包括NQO 1， HO-1、SOD、GPX和Trx。Nrf 2与炎症的关系也进行了探讨。 这种方法将确定每种抗氧化增强策略提供的保护途径， 确定需要进一步研究的先导分子，并继续进行涉及动物毒理学的未来工作， 清除和活动评估。 总之，使用化学分析和生物研究数据的比较方法将使 研究人员鉴定提供作为药物所需特征的分子和部分， 用于治疗由氧化应激引起的神经变性和眼部疾病。这项提案需要一个 通过使用计算和合成化学将联合收割机 将不同的反应性构建块转化为小分子， 神经变性和眼部疾病的分子特征，其方式大于 各个部分。

PROJECT SUMMARY/ABSTRACT Uncontrolled oxidative stress contributes to the development of neurodegenerative and ocular diseases for which there are yet to be effective therapeutic interventions. The investigators have developed small molecules that exhibit significant antioxidant reactivity in biological assays through two mechanisms: direct reactivity with reactive oxygen species and catalytic activation of the Nrf2 pathway. The investigators will incorporate virtual screening, rational design, synthetic chemistry, and molecular biology methods to produce enhanced antioxidant small molecules and understand how the small molecules activate catalytic antioxidant pathways. New molecules produced from this project are expected to provide radical scavenging activity, metabolic stability, and permeability. We propose four mechanisms of achieving improvement in these features compared to parent molecules. Virtual screening will be used to identify what combination of these approaches will yield the most promising targets. Following synthesis of these targets, cellular models for neurodegenerative and ocular disease (e.g. cataracts) will be used to test for protection from oxidative stress. Molecules showing potent antioxidant activity will then be screened for metabolic stability and blood brain barrier permeability. Explorations will also be focused on uncovering the mechanism(s) through which our small molecules protect through catalytic activation of innate antioxidant pathways in biology (Nrf2). Cell lysates from models exposed to ROS will be evaluated for Nrf2 and its downstream genes including NQO1, HO-1, SOD, GPX, and Trx, for example. The relationship between Nrf2 and inflammation is also explored. This approach will identify the path(s) of protection each strategy of antioxidant enhancement provides and identify lead molecules to be explored further and proceed to future work involving animal toxicology, clearance, and activity assessment. Altogether, a comparative approach that uses data from chemical assays and biological studies will allow the investigators to identify molecules and moieties that provide the characteristics needed to serve as a therapeutic for neurodegenerative and ocular disorders arising from oxidative stress. This proposal takes a unique approach to targeting oxidative stress by using computational and synthetic chemistry to combine different reactive building-blocks into small molecules designed to have activity through targeting molecular features of neurodegeneration and diseases of the eye in a manner that is greater than the sum of the individual parts.